Capacitance detection in electrochemical assays

ABSTRACT

A method and system are provided to determine fill sufficiency of a biosensor test chamber by determining capacitance of the test chamber.

This application claims the benefits of priority under 35 USC§119 and/or§120 from prior filed U.S. Provisional Application Ser. No. 61/308,167filed on Feb. 25, 2010, which applications are incorporated by referencein their entirety into this application.

BACKGROUND

Analyte detection in physiological fluids, e.g. blood or blood derivedproducts, is of ever increasing importance to today's society. Analytedetection assays find use in a variety of applications, includingclinical laboratory testing, home testing, etc., where the results ofsuch testing play a prominent role in diagnosis and management in avariety of disease conditions. Analytes of interest include glucose fordiabetes management, cholesterol, and the like. In response to thisgrowing importance of analyte detection, a variety of analyte detectionprotocols and devices for both clinical and home use have beendeveloped.

One type of method that is employed for analyte detection is anelectrochemical method. In such methods, an aqueous liquid sample isplaced into a sample-receiving chamber in an electrochemical cell thatincludes two electrodes, e.g., a counter and working electrode. Theanalyte is allowed to react with a redox reagent to form an oxidizable(or reducible) substance in an amount corresponding to the analyteconcentration. The quantity of the oxidizable (or reducible) substancepresent is then estimated electrochemically and related to the amount ofanalyte present in the initial sample.

Such systems are susceptible to various modes of inefficiency and/orerror. For example, variations in temperatures can affect the results ofthe method. This is especially relevant when the method is carried outin an uncontrolled environment, as is often the case in homeapplications or in third world countries. Errors can also occur when thesample size is insufficient to get an accurate result. Partially filledtest strips can potentially give an inaccurate result because themeasured test currents are proportional to the area of the workingelectrode that is wetted with sample. Thus, partially filled test stripscan under certain conditions provide a glucose concentration that isnegatively biased.

SUMMARY OF THE DISCLOSURE

Applicants believe that effects of parallel strip resistance indetermining filled biosensor test strips have been ignored, leading toinaccurate high measurement of capacitance in a test strip, especiallywhen lower parallel resistance is encountered. Exemplary embodiments ofapplicants' invention take into consideration this effect and at thesame time obviate the need to determine the resistance in a biosensortest chamber.

In one aspect, a method of determining capacitance of a biosensor isprovided. The biosensor includes a chamber having two electrodesdisposed in the chamber and coupled to a microcontroller. The method canbe achieved by: initiating an electrochemical reaction in the biosensorchamber; applying an oscillating voltage of a predetermined frequency tothe chamber; determining a phase angle between a current output and theoscillating voltage from the chamber; and calculating a capacitance ofthe chamber based on a product of the current output and a sine of thephase angle divided by a product of two times pi times the frequency andthe voltage.

In a further aspect, an analyte measurement system is provided thatincludes an analyte test strip and analyte test meter. The analyte teststrip includes a substrate having a reagent disposed thereon, and atleast two electrodes proximate the reagent in test chamber. The analytemeter includes a strip port connector disposed to connect to the twoelectrodes, a power supply, and a microcontroller electrically coupledto the strip port connector and the power supply. The microcontroller isprogrammed to: initiate an electrochemical reaction in the biosensorchamber; apply an oscillating voltage of a predetermined frequency tothe chamber; determine a phase angle between a current output and theoscillating voltage from the chamber; and calculate a capacitance of thechamber based on a product of the current output and a sine of the phaseangle divided by a product of two times pi times the frequency and thevoltage.

In yet another aspect, analyte measurement system is provided thatincludes an analyte test strip and analyte test meter. The test stripincludes a substrate having a reagent disposed thereon, and at least twoelectrodes proximate the reagent in test chamber. The analyte meterincludes a strip port connector disposed to connect to the twoelectrodes, a power supply, and a microcontroller electrically coupledto the strip port connector and the power supply such that a percenterror in capacitance measurement of the test strip across a range ofcapacitance as compared to a referential parallel R-C circuit is lessthan about 3%.

These and other embodiments, features and advantages will becomeapparent to those skilled in the art when taken with reference to thefollowing more detailed description of various exemplary embodiments ofthe invention in conjunction with the accompanying drawings that arefirst briefly described.

BRIEF DESCRIPTION OF THE FIGURES

The accompanying drawings, which are incorporated herein and constitutepart of this specification, illustrate presently preferred embodimentsof the invention, and, together with the general description given aboveand the detailed description given below, serve to explain features ofthe invention (wherein like numerals represent like elements).

FIG. 1 illustrates an exemplary analyte measurement system including ananalyte test meter and test strip.

FIG. 2 illustrates in simplified schematic view of an exemplary circuitboard for the meter of FIG. 1.

FIG. 3 illustrates an exploded perspective view of the test strip ofFIG. 1.

FIG. 4 illustrates a simplified schematic of the components to determinecapacitance of a filled test strip.

FIG. 5A illustrates the application of voltage over time applied to thetest strip.

FIG. 5B illustrates the measured current response from the test stripover time.

FIG. 6A illustrates a sampling of the current output indicated at area602.

FIG. 6B illustrates the alternating current output once thedirect-current component has been removed from the sampled data of FIG.6A.

FIGS. 6C and 6D illustrate the phase angle between the alternatingvoltage applied to the test strip and the alternating current outputfrom the test strip.

FIG. 6E illustrates an interpolation of the sampled data to determinethe cross-over point of FIG. 6D for comparison with the cross-over pointof the applied current of FIG. 6C.

FIG. 7 illustrates an exemplary flow chart of the method to determinecapacitance in the exemplary test strip.

FIG. 8A illustrates the percent error of the exemplary embodimentsversus a known system and other related techniques of the applicants.

FIG. 8B illustrates the distribution of capacitance of respectivecapacitance measurement techniques over the range of resistance in theexemplary test strip.

DETAILED DESCRIPTION OF THE EXEMPLARY FIGURES

The following detailed description should be read with reference to thedrawings, in which like elements in different drawings are identicallynumbered. The drawings, which are not necessarily to scale, depictselected embodiments and are not intended to limit the scope of theinvention. The detailed description illustrates by way of example, notby way of limitation, the principles of the invention. This descriptionwill clearly enable one skilled in the art to make and use theinvention, and describes several embodiments, adaptations, variations,alternatives and uses of the invention, including what is presentlybelieved to be the best mode of carrying out the invention.

As used herein, the terms “about” or “approximately” for any numericalvalues or ranges indicate a suitable dimensional tolerance that allowsthe part or collection of components to function for its intendedpurpose as described herein. In addition, as used herein, the terms“patient,” “host,” “user,” and “subject” refer to any human or animalsubject and are not intended to limit the systems or methods to humanuse, although use of the subject invention in a human patient representsa preferred embodiment.

The subject systems and methods are suitable for use in thedetermination of a wide variety of analytes in a wide variety ofsamples, and are particularly suited for use in the determination ofanalytes in whole blood, plasma, serum, interstitial fluid, orderivatives thereof. In an exemplary embodiment, a glucose test systembased on a thin-layer cell design with opposing electrodes and tri-pulseelectrochemical detection that is fast (e.g., about 5 second analysistime), requires a small sample (e.g., about 0.4 μL(microliter)), and canprovide improved reliability and accuracy of blood glucose measurements.In the reaction cell, glucose in the sample can be oxidized togluconolactone using glucose dehydrogenase and an electrochemicallyactive mediator can be used to shuttle electrons from the enzyme to aworking electrode. A potentiostat can be utilized to apply a tri-pulsepotential waveform to the working and counter electrodes, resulting intest current transients used to calculate the glucose concentration.Further, additional information gained from the test current transientsmay be used to discriminate between sample matrices and correct forvariability in blood samples due to hematocrit, temperature variation,electrochemically active components, and identify possible systemerrors.

The subject methods can be used, in principle, with any type ofelectrochemical cell having spaced apart first and second electrodes anda reagent layer. For example, an electrochemical cell can be in the formof a test strip. In one aspect, the test strip may include two opposingelectrodes separated by a thin spacer for defining a sample-receivingchamber or zone in which a reagent layer is located. One skilled in theart will appreciate that other types of test strips, including, forexample, test strips with co-planar electrodes may also be used with themethods described herein.

FIG. 1 illustrates a diabetes management system that includes a diabetesdata management unit 10 and a biosensor in the form of a glucose teststrip 80. Note that the diabetes data management unit (DMU) may bereferred to as an analyte measurement and management unit, a glucosemeter, a meter, and an analyte measurement device. In an embodiment, theDMU may be combined with an insulin delivery device, an additionalanalyte testing device, and a drug delivery device. The DMU may beconnected to the computer 26 or server 70 via a cable or a suitablewireless technology such as, for example, GSM, CDMA, BlueTooth, WiFi andthe like.

Referring back to FIG. 1, glucose meter 10 can include a housing 11,user interface buttons (16, 18, and 20), a display 14, and a strip portopening 22. User interface buttons (16, 18, and 20) can be configured toallow the entry of data, navigation of menus, and execution of commands.User interface button 18 can be in the form of a two way toggle switch.Data can include values representative of analyte concentration, and/orinformation, which are related to the everyday lifestyle of anindividual. Information, which is related to the everyday lifestyle, caninclude food intake, medication use, occurrence of health check-ups, andgeneral health condition and exercise levels of an individual.

The electronic components of meter 10 can be disposed on a circuit board34 that is within housing 11. FIG. 2 illustrates (in simplifiedschematic form) the electronic components disposed on a top surface ofcircuit board 34. On the top surface, the electronic components mayinclude a strip port opening 308, a microcontroller 38, a non-volatileflash memory 306, a data port 13, a real time clock 42, and a pluralityof operational amplifiers (46-49). On the bottom surface, the electroniccomponents may include a plurality of analog switches, a backlightdriver, and an electrically erasable programmable read-only memory(EEPROM, not shown). Microcontroller 38 can be electrically connected tostrip port opening 308, non-volatile flash memory 306, data port 13,real time clock 42, the plurality of operational amplifiers (46-49), theplurality of analog switches, the backlight driver, and the EEPROM.

Referring back to FIG. 2, the plurality of operational amplifiers caninclude gain stage operational amplifiers (46 and 47), a trans-impedanceoperational amplifier 48, and a bias driver operational amplifier 49.The plurality of operational amplifiers can be configured to provide aportion of the potentiostat function and the current measurementfunction. The potentiostat function can refer to the application of atest voltage between at least two electrodes of a test strip. Thecurrent function can refer to the measurement of a test currentresulting from the applied test voltage. The current measurement may beperformed with a current-to-voltage converter. Microcontroller 38 can bein the form of a mixed signal microprocessor (MSP) such as, for example,the Texas Instrument MSP 430. The MSP 430 can be configured to alsoperform a portion of the potentiostat function and the currentmeasurement function. In addition, the MSP 430 can also include volatileand non-volatile memory. In another embodiment, many of the electroniccomponents can be integrated with the microcontroller in the form of anapplication specific integrated circuit (ASIC).

Strip port connector 308 can be located proximate the strip port opening22 and configured to form an electrical connection to the test strip.Display 14 can be in the form of a liquid crystal display for reportingmeasured glucose levels, and for facilitating entry of lifestyle relatedinformation. Display 14 can optionally include a backlight. Data port 13can accept a suitable connector attached to a connecting lead, therebyallowing glucose meter 10 to be linked to an external device such as apersonal computer. Data port 13 can be any port that allows fortransmission of data such as, for example, a serial, USB, or a parallelport.

Real time clock 42 can be configured to keep current time related to thegeographic region in which the user is located and also for measuringtime. Real time clock 42 may include a clock circuit 45, a crystal 44,and a super capacitor 43. The DMU can be configured to be electricallyconnected to a power supply such as, for example, a battery. The supercapacitor 43 can be configured to provide power for a prolonged periodof time to power real time clock 42 in case there is an interruption inthe power supply. Thus, when a battery discharges or is replaced, realtime clock does not have to be re-set by the user to a proper time. Theuse of real time clock 42 with super capacitor 43 can mitigate the riskthat a user may re-set real time clock 42 incorrectly.

FIG. 3 illustrates an exemplary test strip 80, which includes anelongate body extending from a distal end 80 to a proximal end 82, andhaving lateral edges. As shown here, the test strip 80 also includes afirst electrode layer 66 a, insulation layer 66 b, a second electrodelayer 64 a, insulation layer 64 b, and a spacer 60 sandwiched in betweenthe two electrode layers 64 a and 66 a. The first electrode layer 66 acan include a first electrode 67 a, a first connection track 76, and afirst contact pad 47, where the first connection track 76 electricallyconnects the first electrode layer 66 a to the first contact pad 67, asshown in FIGS. 3 and 4. Note that the first electrode 67 a is a portionof the first electrode layer 66 a that is immediately underneath thereagent layer 72. Similarly, the second electrode layer 64 a can includea second electrode 67 b, a second connection track 78, and a secondcontact pad 78, where the second connection track 78 electricallyconnects the second electrode 67 b with the second contact pad 78, asshown in FIGS. 3 and 4. Note that the second electrode includes aportion of the second electrode layer 64 a that is above the reagentlayer 72.

As shown in FIG. 3, the sample-receiving chamber 61 is defined by thefirst electrode, the second electrode, and the spacer 60 near the distalend 80 of the test strip 80. The first electrode 67 a and the secondelectrode 67 b can define the bottom and the top of sample-receivingchamber 61, respectively. A cutout area 68 of the spacer 60 can definethe sidewalls of the sample-receiving chamber 61. In one aspect, thesample-receiving chamber 61 can include ports 70 that provide a sampleinlet and/or a vent. For example, one of the ports can allow a fluidsample to ingress and the other port can allow air to egress. In oneexemplary embodiment, the first electrode layer 66 a and the secondelectrode layer 64 a can be made from sputtered palladium and sputteredgold, respectively. Suitable materials that can be employed as spacer 60include a variety of insulating materials, such as, for example,plastics (e.g., PET, PETG, polyimide, polycarbonate, polystyrene),silicon, ceramic, glass, adhesives, and combinations thereof. In oneembodiment, the spacer 60 may be in the form of a double sided adhesivecoated on opposing sides of a polyester sheet where the adhesive may bepressure sensitive or heat activated.

Referring back to FIG. 3, the area of first electrode and secondelectrode can be defined by the two lateral edges and cutout area 68.Note that the area can be defined as the surface of the electrode layerthat is wetted by liquid sample. In an embodiment, the adhesive portionof spacer 60 can intermingle and/or partially dissolve the reagent layerso that the adhesive forms a bond to the first electrode layer 66A. Suchan adhesive bond helps define the portion of the electrode layer thatcan be wetted by liquid sample and also electrooxidize or electroreducemediator.

Either the first electrode or the second electrode can perform thefunction of a working electrode depending on the magnitude and/orpolarity of the applied test voltage. The working electrode may measurea limiting test current that is proportional to the reduced mediatorconcentration. For example, if the current limiting species is a reducedmediator (e.g., ferrocyanide), then it can be oxidized at the firstelectrode as long as the test voltage is sufficiently less than theredox mediator potential with respect to the second electrode. In such asituation, the first electrode performs the function of the workingelectrode and the second electrode performs the function of acounter/reference electrode. Note that one skilled in the art may referto a counter/reference electrode simply as a reference electrode or acounter electrode. A limiting oxidation occurs when all reduced mediatorhas been depleted at the working electrode surface such that themeasured oxidation current is proportional to the flux of reducedmediator diffusing from the bulk solution towards the working electrodesurface. The term bulk solution refers to a portion of the solutionsufficiently far away from the working electrode where the reducedmediator is not located within a depletion zone. It should be noted thatunless otherwise stated for test strip 80, all potentials applied bytest meter 10 will hereinafter be stated with respect to secondelectrode. Similarly, if the test voltage is sufficiently greater thanthe redox mediator potential, then the reduced mediator can be oxidizedat the second electrode as a limiting current. In such a situation, thesecond electrode performs the function of the working electrode and thefirst electrode performs the function of the counter/referenceelectrode. Details regarding the exemplary test strip, operation of thestrip and the test meter are found in U.S. Patent ApplicationPublication No. 20090301899, which is incorporated by reference in itsentirety herein, with a copy attached to the Appendix.

Referring to FIG. 3, test strip 80 can include one or more workingelectrodes and a counter electrode. Test strip 80 can also include aplurality of electrical contact pads, where each electrode can be inelectrical communication with at least one electrical contact pad. Stripport connector 308 can be configured to electrically interface to theelectrical contact pads and form electrical communication with theelectrodes. Test strip 80 can include a reagent layer that is disposedover at least one electrode. The reagent layer can include an enzyme anda mediator. Exemplary enzymes suitable for use in the reagent layerinclude glucose oxidase, glucose dehydrogenase (with pyrroloquinolinequinone co-factor, “PQQ”), and glucose dehydrogenase (with flavinadenine dinucleotide co-factor, “FAD”). An exemplary mediator suitablefor use in the reagent layer includes ferricyanide, which in this caseis in the oxidized form. The reagent layer can be configured tophysically transform glucose into an enzymatic by-product and in theprocess generate an amount of reduced mediator (e.g., ferrocyanide) thatis proportional to the glucose concentration. The working electrode canthen measure a concentration of the reduced mediator in the form of acurrent. In turn, glucose meter 10 can convert the current magnitudeinto a glucose concentration. Details of the preferred test strip areprovided in U.S. Pat. Nos. 6,179,979; 6,193,873; 6,284,125; 6,413,410;6,475,372; 6,716,577; 6,749,887; 6,863,801; 6,890,421; 7,045,046;7,291,256; 7,498,132, all of which are incorporated by reference intheir entireties herein.

FIG. 4 illustrates, in simplified schematic form, of various functionalcomponents utilized for capacitance determination. In particular, thecomponents include a microcontroller 300. A preferred embodiment of themicrocontroller 300 is available from Texas Instrument as ultra-lowpower microcontroller model MSP430. Microcontroller (“MC”) 300 may beprovided with DAC output and built-in A-D conversion. MC 300 is suitablyconnected to a LCD screen 304 to provide a display of the test resultsor other information related to the test results. Memory 306 iselectrically connected to the MC 300 for storage of test results, sensedcurrent and other necessary information or data. The test strip may becoupled for a test measurement via a strip port connector (“SPC”) 308.SPC 308 allows the test strip to interface with MC 300 via a firstcontact pad 47 a, 47 b and a second contact pad 43. The second contactpad 43 can be used to establish an electrical connection to the testmeter through a U-shaped notch 45, as illustrated in FIG. 4. SPC 308 mayalso be provided with electrode connectors 308 a and 308 c. The firstcontact pad 47 can include two prongs denoted as 47 a and 47 b. In oneexemplary embodiment, the first electrode connectors 308 a and 308 cseparately connect to prongs 47 a and 47 b, respectively. The secondelectrode connector 308 b can connect to second contact pad 43. The testmeter 10 can measure the resistance or electrical continuity between theprongs 47 a and 47 b to determine whether the test strip 80 iselectrically connected to the test meter 10.

Referring to FIG. 4, SPC 308 is connected to switch 310. Switch 310 isconnected to the bias driver 312. Bias driver 312 is provided with theDAC signal 312 a; current drive 312 b and switch signal 312 c. The MC300 provides the DAC signal 312 a, which includes analogue voltages inthe range 0 to Vref (e.g., about 2.048V). The bias driver 312 canoperate in two modes—constant voltage, or constant current. Thecurrent-driver line 312 b controls the mode of the bias driver 312.Setting the line 312 b low converts an op-amp in the bias driver 312 toa voltage follower amplifier. DAC signal 312 a output is scaled toVref/2+/−400 mV full scale. The op-amp in the bias driver outputs thisvoltage directly to the MC 300 as line driver-line 312 d. The voltage ofline 312 d is generated with respect to the Vref/2 virtual ground. So todrive a suitable bias (e.g., about 20 mV bias), the DAC must drive(through a suitable scaler) about 1.044V. To drive a bias of about +300mV, the DAC must generally provide about 1.324V, and for the −300 mVbias, the DAC must generally provide about 0.724V. The bias drivercircuit 312 also generates the 109 Hz sine wave, which is used for filldetection via capacitance measurement.

On the other hand, if current-drive signal 312 a to bias driver 312 isheld high, the DAC output is scaled to approximately 0 to approximately60 mV full scale. Switch signal 312 c may also be energized, causing thecurrent path through the test strip to be diverted through a resistor inbias driver 312. The op-amp in bias driver 312 attempts to control thevoltage drop across the resistor to be the same as the scaled DACdrive—producing in this case a current of approximately 600 nA. Thiscurrent is used for sample detection in order to initiate a testmeasurement.

Bias driver 312 is also connected to a trans-impedance amplifier circuit(“TIA circuit”) 314. TIA circuit 314 converts the current flowing thoughthe strip's electrode layer 66 a (e.g., palladium) to electrode layer 64a (e.g., gold) contacts into a voltage. The overall gain is controlledby a resistor in TIA circuit 314. Because the strip 80 is a highlycapacitive load, normal low-offset amplifiers tend to oscillate. Forthis reason a low-cost op-amp is provided in the TIA circuit 314 as aunity gain buffer and incorporated within the overall feedback loop. Asa functional block, circuit 314 acts as dual op-amp system with bothhigh drive capability and low voltage offset. The TIA circuit 314 alsoutilizes a virtual ground (or virtual earth) to generate the 1.024V biason the electrode layer 64 a (e.g., gold) contact of the SPC 308. Circuit314 is also connected to a Vref amplifier circuit 316. This circuit,when in current measuring mode, uses a virtual ground rail set at Vref/2(approximately 1.024V), allowing both positive and negative currents tobe measured. This voltage feeds all of the gain amplifier stage 318. Toprevent any circuit loads from ‘pulling’ this voltage, a unity gainbuffer amplifier may be utilized within the Vref amplifier circuit 316.

The strip current signal 314 a from the TIA circuit 314 and the virtualground rail 316 a (˜Vref/2) from the voltage reference amplifier 316 arescaled up as needed for various stages of the test measurement cycle. Inthe exemplary embodiment, MC 300 is provided with four channels ofamplified signal sensed from the test strip with varying amplificationsof the sensed current as need for different stages of the measurementcycle of the test strip during an analyte assay.

In one embodiment, the test meter 10 can apply a test voltage and/or acurrent between the first contact pad 47 and the second contact pad 43of the test strip 80. Once the test meter 10 recognizes that the strip80 has been inserted, the test meter 10 turns on and initiates a fluiddetection mode. In one embodiment, the meter attempts to drive a smallcurrent (e.g. 0.2 to 1 μA) through the strip 80. When there is no samplepresent the resistance is greater than several Mega Ohms, so the drivingvoltage on the op-amp trying to apply the current goes to the rail. Whena sample is introduced the resistance drops precipitously and thedriving voltage follows. When the driving voltage drops below apre-determined threshold the test sequence is initiated.

FIG. 5A shows the voltage to be applied between the electrodes. Timezero is taken to be when the sample detection method has detected that asample first begins to fill the strip. Note that the sine wave componentshown at approximately 1.3 seconds in FIG. 5A is not drawn on thecorrect timescale for illustration purposes.

After a sample has been detected in the test strip chamber 61, thevoltage between the strip electrodes is stepped to a suitable voltage inmillivolts of magnitude and maintained for a set amount of time, e.g.,about 1 second, then stepped to a higher voltage and held for a fixedamount of time, then a sine wave voltage is applied on top of the DCvoltage for a set amount of time, then the DC voltage is applied for afurther amount of time, then reversed to a negative voltage and held fora set amount of time. The voltage is then disconnected from the strip.This series of applied voltages generates a current transient such asthe one shown in FIG. 5B.

In FIG. 5B, the current signal from about 0 to about 1 second (as wellas later current samples) may be used for error checking and todistinguish a control solution sample from a blood sample. The signalfrom about 1 to about 5 seconds is analyzed to obtain a glucose result.The signal during this period is also analyzed for various errors. Thesignal from about 1.3 to 1.4 seconds is used to detect whether or notthe sensor is completely filled with sample. The current from 1.3 to1.32 seconds, denoted here as trace 500, is sampled at approximately 150microsecond intervals to determine whether sufficient volume ofphysiological fluid has filled chamber 61 of the test strip.

In one embodiment for performing a sufficient volume check, acapacitance measurement is used to infer sufficient analyte fill of thechamber 61 of the test strip 80. A magnitude of the capacitance can beproportional to the area of an electrode that has been coated withsample fluid. Once the magnitude of the capacitance is measured, if thevalue is greater than a threshold and thus the test strip has asufficient volume of liquid for an accurate measurement, a glucoseconcentration can be outputted. But if the value is not greater than athreshold, indicating that the test strip has insufficient volume ofliquid for an accurate measurement, and then an error message can beoutputted.

In one method for measuring capacitance, a test voltage having aconstant component and an oscillating component is applied to the teststrip. In such an instance, the resulting test current can bemathematically processed, as described in further detail below, todetermine a capacitance value.

Applicants believe that the biosensor test chamber 61 with the electrodelayers can be modeled in the form of a circuit having a parallelresistor and capacitor as shown in Table 1.

In this model in Table 1, R represents the resistance encountered by thecurrent and C represents a capacitance resulting from the combination ofthe physiological fluid and reagent electrically coupled to theelectrodes. To initiate a determination of capacitance of the chamber,an alternating bias voltage may be applied across the respectiveelectrodes disposed in the chamber, and a current from the chamber ismeasured. The filling of the chamber 61 is believed to be generally ameasure of capacitance only and thus any parasitic resistance, such as,for example, R, must not be included in any determination or calculationof capacitance. Hence, in measuring or sensing the current, anyparasitic resistance is believed to affect the measured current.Applicants, however, have discovered a technique to derive capacitancewithout requiring utilization or knowledge of the resistance through thechamber as modeled above. In order to further explain this technique, ashort discussion of the mathematical foundation underlying the techniqueis warranted.

According to Kirchhoff's Law, total current (i_(T)) through the circuitof Table 1 is approximately the sum of the current flowing through theresistor (i_(R)) and through the capacitor (i_(C)). When an alternatingvoltage V (as measured as RMS) is applied, the resistor current (i_(R))may be expressed as:

i _(R) =V/R  Eq. 1

Capacitor current (i_(C)) can be expressed as:

i _(C) =jωCV  Eq. 2

-   -   Where:        -   j is an imaginary number operator indicating that current            leads voltage by about 90 degrees in a capacitor; and        -   ω is the angular frequency 2πƒ where f is frequency in            Hertz.

The summation of these components is shown in the phasor diagram ofTable 1. In the phasor diagram, Φ represents the phase angle of theinput as compared to the output. Phase angle Φ is determined by thefollowing trigonometric function:

tanΦ=I _(c)/I _(R)  Eq. 3

By Pythagoras theorem, the square of the total current i_(T) can becalculated as:

i _(T) ² =i _(C) ² +i _(R) ²  Eq. 4

By rearranging Eq. 4 and substituting Eq. 3, the following equation isarrived at:

i _(C) ² =i _(T) ²−^(i) ^(C) ²/_((tanΦ)) ²  Eq. 5

Resolving for capacitor current i_(C) and combining with Eq. 2:

i _(C)=√{square root over (()}i _(T) ²*(tanΦ)²/((tan Φ))²+1))=ωCV  Eq. 6

Rearranging for C and expanding ω, the capacitance becomes:

C=(√{square root over (()}i _(T) ²*(tan Φ)²/((tan Φ)²+1))/2πƒV  Eq. 7

Simplification of Eq. 7 leads to:

C=|(i _(T)sinΦ)|/2πƒV  Eq. 8

It can be seen that Eq. 8 does not reference to the resistor current.Consequently, if the system can drive an alternating voltage withfrequency f and root-mean-squared (“RMS”) amplitude V, and measure totalcurrent i_(T) as RMS value and phase angle Φ, capacitance C of the testchamber 61 can be accurately calculated without having to determineresistance in the biosensor test chamber. This is believed to be ofsubstantial benefit because the resistance of the biosensor strip isdifficult to measure, and varies over the 5 second assay time.Resistance is believed to arise from how many charge carriers can flowthrough the strip for a given electrical bias (voltage), and istherefore reaction dependent. At the 1.3 second point in the assay, theresistance is expected to be anything from 10 kΩ to perhaps 100 kΩ.Hence, by not having to determine the resistance in the biosensorchamber or even the resistance in the measuring circuit, such as asensor resistor, applicants' invention have advanced the state of theart in improving of the entire test strip.

Implementation of an exemplary technique to determine capacitance Cbased on Eq. 8 can be understood in relation FIGS. 6A, 6B, 6C, 6D, 6E,and 7. As illustrated in FIG. 5A and step 702 of FIG. 7, an AC testvoltage (.±0.50 mV peak-to-peak) of approximately 109 Hz can be appliedfor 2 cycles during approximately 1-1.3 seconds or at least one cycleindicated in step 704. In the preferred embodiments, the first cycle canbe used as a conditioning pulse and the second cycle can be used todetermine the capacitance. The alternating test voltage can be of asuitable waveform, such as, for example, a sine wave of approximately109 Hertz with approximately 50 millivolts peak (FIG. 6C). The samplingcan be of any suitable sampling size per cycle, such as, for exampleapproximately 64-65 samples per cycle, shown here in FIG. 6A. Hence,each sample represents approximately 5.6 degrees of the exemplary sinewave.

In FIG. 6A, the system adds a direct-current voltage offset to thealternating current bias and therefore the measured samples in FIG. 6Awill also have a direct-current offset, which must be removed via steps706 and 708 in order to determine the total current i_(T) according toone example of applicant's technique.

In this technique, a mean of all the 65 samples, referenced here as 602,in FIG. 6A is derived in step 706, which will provide a threshold forthe zero current of the a.c. component of the samples. A benefit of thisderivation is that the noise across the samples is averaged out. Foreach sample point, the mean value is subtracted out of each sampledpoint in step 708, which results in isolating the alternating currentcomponent, shown here in FIG. 6B. Thereafter, a RMS value of all thenegative values is taken in step 710, to provide for a substantiallyaccurate magnitude of the total current i_(T). It is noted that the RMSvalue of the positive values could also be taken, but applicants believethat the positive values are disjointed due to being split across thefirst and fourth quadrants of the overall cycle, and therefore thenegative values are preferred. Once the samples 602 have beenmanipulated to remove the DC offset, the samples can be plotted to showthe output of the current over time, as referenced here at 604 in FIG.6B.

To determine the phase angle, the system or MC, as appropriatelyprogrammed can compare the oscillating input voltage, shown here in FIG.6C to the oscillating output current to determine the phase angle forstep 714. In the preferred embodiments, the sampled data 604 is analyzedto determine a cross-over point from positive to negative current.Because the sampling is based on a discrete number of samples,interpolation can be used to determine substantially when the outputcurrent crosses over the zero current line in FIG. 6E, the interpolatedcross-over point being referenced here as 608. In the embodimentdescribed here, the phase angle Φ is less than 90 degrees andapproximately 87 degrees. For increased accuracy, interpolation can beperformed at another cross-over point 610 with approximately 180 degreessubtracted from this second interpolated point 610. The two interpolatedvalues should be within a few degrees and may be averaged out toincrease accuracy.

Once the phase angle has been derived, capacitance can be calculatedusing Eq. 8. In practice, however, it has been determined that theimplementation of the trans-impedance amplifier 314 and the gainamplifier introduces additional phase shift into the system. Thisadditional phase shift can be offset by introduction of a compensationvalue Φ_(COMP) by measuring the capacitance of the system without astrip in use.

C=|i _(T)sin(Φ+Φ_(COMP))|/2πƒV  Eq. 9

In the preferred embodiments, the compensation phase angle Φ_(COMP)ranges from about 5 to about 7 degrees.

Once capacitance of the test strip 80 has been determined, a two-pointcalibration can be performed to normalize the capacitance value to avalue that is independent of any tolerances of the analog components(e.g., resistors, capacitors, op-amps, switches and the like). Briefly,the two-point calibration is performed by: placing a 550 nF capacitorwith 30 k parallel resistance across the measurement input; command themeter to measure the capacitance, and note the value produced; place a800 nF capacitor with 30 k parallel resistance across the measurementinput; command the meter to measure the capacitance, and note the valueproduced. These two points will give an indication of the gain andoffset of the measurement capability of that particular hardwareinstance (not the design). A slope and offset are then calculated fromthe measurement errors, and stored in the meter's memory. The meter isnow calibrated.

-   When a strip is inserted and a sample applied, the capacitance is    measured and the stored slope and offset are applied to correct the    measurement.

After completion of the device calibration, an evaluation is made todetermine whether the test chamber 61 has been sufficiently filled withtest fluid. The evaluation can be based on a capacitance magnitude of atleast 65% to 85% of an average capacitance value derived from a largesample of good filled test strips.

To test the robustness of this exemplary technique, applicantsintentionally introduced noise into the system to determine the percenterror as compared to referential parallel R-C circuit. In Table 2 below,despite the number of Analog-to-Digital-Converter (“ADC”) noise countswere introduced, error relating to current, phase angle and capacitancewere less than 1%.

TABLE 2 ADC Noise Current Phase Angle Capacitance Counts Error (%) Error(%) Error (%) ±1 −0.05 −0.1 −0.09 ±2 −0.08 −0.19 −0.21 ±3 0.2 −0.34−0.34 ±4 0.21 0.39 0.37

Comparison of the exemplary techniques with other techniques confirmsthe increased accuracy of applicants' technique. For example, in FIG.8A, capacitance is measured from a sample of strips in the range ofabout 350 to about 800 nanoFarad. A fully filled strip has capacitanceranging between 600 and 700 nF depending on whether control solution orblood is used. Partially filled strips exhibit lower capacitance ofcourse. The capacitance is measured with the subject embodiment todetermine percent deviation from a referential parallel R-C circuit. Thepercentage error is calculated by having several “golden” R-Ccombinations that have been calibrated using a commercially availableLCR meter. These R-C combinations (which have been found as generallyerror-free exemplars and therefore are “golden”) are presented to thestrip connector in turn, and the system is commanded to read thecapacitance. This test is repeated using several other samples of thesystem to determine the precision and reliability of the measurementtechnique. Reference curve 800 represents the exemplary technique witherror rate from the referential datum of less than 3% through thecapacitance range of about 350 nanoFarad to about 850 nanoFarads. Incontrast, capacitance measurement in an existing meter system availablefrom LifeScan Inc., in the Netherlands shows error curve 806 rangingfrom less than 2 percent to greater than 10 percent through this rangeof capacitance. Applicants' related capacitance measurement techniques802 and 804 fall in between the upper boundary 806 sets by the existinganalyte measurement system and the lower boundary 800 sets by theexemplary technique.

Although the exemplary embodiments, methods, and system have beendescribed in relation to a blood glucose strip, the principles describedherein are also applicable to any analyte measurement strips thatutilize a physiological fluid on a reagent disposed between at least twoelectrodes.

As noted earlier, the microcontroller can be programmed to generallycarry out the steps of various processes described herein. Themicrocontroller can be part of a particular device, such as, forexample, a glucose meter, an insulin pen, an insulin pump, a server, amobile phone, personal computer, or mobile hand held device.Furthermore, the various methods described herein can be used togenerate software codes using off-the-shelf software development toolssuch as, for example, C or variants of C such as, for example, C+, C++,or C-Sharp. The methods, however, may be transformed into other softwarelanguages depending on the requirements and the availability of newsoftware languages for coding the methods. Additionally, the variousmethods described, once transformed into suitable software codes, may beembodied in any computer-readable storage medium that, when executed bya suitable microcontroller or computer, are operable to carry out thesteps described in these methods along with any other necessary steps.

While the invention has been described in terms of particular variationsand illustrative figures, those of ordinary skill in the art willrecognize that the invention is not limited to the variations or figuresdescribed. In addition, where methods and steps described above indicatecertain events occurring in certain order, those of ordinary skill inthe art will recognize that the ordering of certain steps may bemodified and that such modifications are in accordance with thevariations of the invention. Additionally, certain of the steps may beperformed concurrently in a parallel process when possible, as well asperformed sequentially as described above. Therefore, to the extentthere are variations of the invention, which are within the spirit ofthe disclosure or equivalent to the inventions found in the claims, itis the intent that this patent will cover those variations as well.

1. A method of determining capacitance of a biosensor chamber having atwo electrodes disposed in the chamber and coupled to a microcontroller,the method comprising: initiating an electrochemical reaction in thebiosensor chamber; applying an oscillating voltage of a predeterminedfrequency to the chamber; determining a phase angle between a currentoutput and the oscillating voltage from the chamber; and calculating acapacitance of the chamber based on a product of the current output anda sine of the phase angle divided by a product of two times pi times thefrequency and the voltage.
 2. The method of claim 1, in which thecalculating comprises calculating capacitance with an equation of theform:C=|(i _(T)sinΦ)|÷2πƒV where: C≈capacitance; i_(T)≈total current; Φ≈phaseangle between total current and resistor current; ƒ≈frequency; andV≈voltage.
 3. The method of claim 2, in which the calculating comprises:sampling a plurality of current outputs from the chamber over one cycleof the frequency; obtaining a mean of sampled current output;subtracting the mean from each sampled current of the plurality ofcurrent outputs; and extracting root-mean-squared value of all negativevalues from the subtracting to provide for the total current output. 4.The method of claim 3, in which the calculating comprises: determiningfrom the sampling, at least one cross-over point of the current fromnegative to positive values; and interpolating proximate the at leastone cross-over point of the current to determine a first angle at whichthe current changes from positive to negative or negative to positive.5. The method of claim 4, in which the interpolating the at least onecross-over point of the current comprises: interpolating anothercross-over point from the sampling to determine another angle at whichthe current changes from positive to negative or negative to positive;and subtracting from the another angle approximately 180 degrees toprovide for a second angle.
 6. The method of claim 5, in which thesubtracting further comprises calculating an average of the first andsecond angles.
 7. The method of claim 5, in which the calculatingcomprises determining a difference in the angle between the oscillatinginput current and the output current as the phase angle.
 8. An analytemeasurement system comprising: An analyte test strip including: asubstrate having a reagent disposed thereon; at least two electrodesproximate the reagent in test chamber; an analyte meter including: astrip port connector disposed to connect to the two electrodes; a powersupply; and a microcontroller electrically coupled to the strip portconnector and the power supply, the microcontroller being programmed to:(a) initiate an electrochemical reaction in the biosensor chamber; applyan oscillating voltage of a predetermined frequency to the chamber; (b)determine a phase angle between a current output and the oscillatingvoltage from the chamber; and (c) calculate a capacitance of the chamberbased on a product of the current output and a sine of the phase angledivided by a product of two times pi times the frequency and thevoltage.
 9. An analyte measurement system comprising: An analyte teststrip including: a substrate having a reagent disposed thereon; at leasttwo electrodes proximate the reagent in test chamber; an analyte meterincluding: a strip port connector disposed to connect to the twoelectrodes; a power supply; and a microcontroller electrically coupledto the strip port connector and the power supply such that a percenterror in capacitance measurement of the test strip across a range ofcapacitance as compared to a referential parallel R-C circuit is lessthan about 3%.